Cloning of mouse 4ª²1BBL gene and its expression in COSª²7 cells

    LIU Chengª²Li1, DOU Keª²Feng1, ZHU Bangª²Fu2, CAO Yunª²Xin3, ZANG Xiaoª²Xia4, CHAI Yuª²Bo2, DU Keª²Jun2, CHEN Suª²Min2

    1Department of Hepatobiliary Surgery, Xijing Hospital, 2Department of Biochemistry, 3Department of Immunology, School of Basic Medicine, 4Department of Odental Diseases, Stomatological College, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To obtain mouse 4ª²1BBL gene and construct its eukaryotic expression vector and express it in mammalian cells. METHODS: RTª²PCR was applied to amplify mouse 4ª²1BBL gene from total RNA of C57BL/6 mouse spleen cells after stimulating by PHA. Then cloning vector of m4ª²1BBL was constructed by gene recombination technique. After sequencing, the cDNA was recombinated into eukaryotic expression vector pCDNA3.1(+) and transfected into COSª²7 cells. m4ª²1BBLmRNA of transfected cells was detected by RTª²PCR. Indirect immunofluorescent methods and flow cytometry were applied to test the expression of m4ª²1BBL protein. RESULTS: The m4ª²1BBL cDNA was obtained from C57BL/6 mouse spleen cells and was confirmed correct by sequence analysis. The COSª²7 cells transfected with pCDNA3.1(+)ª²m4ª²1BBL could efficiently express m4ª²1BBL mRNA and protein. CONCLUSION: The cloning of mouse 4ª²1BBL gene and the construction of its eukaryotic expression vector were all successful. This study lay the foundation for further research on m4ª²1BBL. ......